Differential dielectroscopic data on the relation of erythrocyte membrane skeleton to erythrocyte deformability and...

Two dielectric relaxations, β_sp (1.5 MHz) and γ1_sp (7 MHz), have been detected on spectrin-based membrane skeleton (MS) of red blood cells (RBCs) using the plot of admittance changes at the spectrin denaturation temperature (Ivanov and Paarvanova in Bioelectrochemistry 110: 59-68, 2016, Electrochim Acta 317: 289-300, 2019a). In this study, we treated RBCs and RBC ghost membranes with agents that make membranes rigid and suppress membrane flicker, and studied the effect on β_sp and γ1_sp relaxations. Diamide (diazene dicarboxylic acid bis-(N,N-dimethylamide)) (up to 0.85 mM), taurine mustard (tris(2-chloroethyl)amine) (up to 2 mM), known to specifically cross-link and stiffen spectrin, and glutaraldehyde (up to 0.044%) all inhibited the relaxations in RBC ghost membranes. Similar inhibition was obtained resealing RBC ghost membranes with 2,3-diphosphoglicerate (up to 15 mM), binding WGA (wheat germ agglutinin) (up to 0.025 mg/ml) to exofacial aspect of RBCs, incubating RBCs in hypotonic (200 mOsm) and hypertonic (600-900 mOsm) media and depleting RBCs of ATP. By contrast, concanavalin A (1 mg/ml) and DIDS (4,4'-diiso-thiocyanato stilbene-2,2'-disulfonic acid) (75 μM, pH 8.2), both known to bind specifically band 3 integral protein of RBCs without effect on RBC membrane rigidity, did not affect the relaxations. We conclude there might be a relation between the strength of dielectric relaxations on MS spectrin and the deformability and flicker of RBC membrane.